Analysis of human α-thrombin by hydrophobic interaction high-performance liquid chromatography

Karlsson, Göran

doi:10.1016/s1046-5928(02)00596-x

Cited by 5 publications

(2 citation statements)

References 13 publications

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“…thrombin with similar efficiency but with significantly lower risk of immunologic impact (Chapman et al, 2007) (Chang, 1986), sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE), hydrophobic interaction HPLC (Karlsson, 2003). Presented work has been focused on development of analytical method based on RP-HPLC.…”

Section: Comparison Of Two Rp-hplc Methods For Determination Of Recommentioning

confidence: 99%

Comparison of two RP-HPLC methods for determination of recombinant human thrombin in pharmaceutical formulations. / Porovnanie dvoch RP-HPLC metód pre stanovenie rekombinantného ľudského trombínu vo farmaceutických formuláciách

Bartoš

Murányi

Šnauko

2015

Acta Facultatis Pharmaceuticae Universitatis Comenianae

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Two reversed-phase high performance liquid chromatography analytical methods (Method I and Method II) for determination of assay of recombinant human thrombin in pharmaceutical formulations were developed and validated. Analysis was performed on chromatographic system Agilent 1200 series SL with diode array detection and mass selective detection. Method I was intended for faster determination of thrombin assay. Gradient programme was optimised to achieve sufficient separation and acceptable runtime. Chromatographic analysis was performed on analytical column Grace Vydac, C4 250 × 4.6 mm, 5 mm. Method II is Method I adapted to use the mass selective detector. Chromatographic separation was performed on analytical column Zorbax 300SB-C8 SolvSaver Plus, 150 × 3 mm, 3.5 mm. Both analytical methods were validated with respect to specificity, linearity, precision and accuracy. The response of thrombin was a linear function of concentration over the range 0.1-1.0 mg/ml. Precision and accuracy of thrombin was evaluated at three concentration levels low (0.2 mg/ml), medium (0.4 mg/ml) and high (0.8 mg/ml). Both validated methods have been successfully applied for determination of assay and thrombin degradation products in pharmaceutical formulations.

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Section: Comparison Of Two Rp-hplc Methods For Determination Of Recommentioning

confidence: 99%

Comparison of two RP-HPLC methods for determination of recombinant human thrombin in pharmaceutical formulations. / Porovnanie dvoch RP-HPLC metód pre stanovenie rekombinantného ľudského trombínu vo farmaceutických formuláciách

Bartoš

Murányi

Šnauko

2015

Acta Facultatis Pharmaceuticae Universitatis Comenianae

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“…Hydrophobic interaction chromatography (HIC) is now widely used as a method for the purification of monoclonal antibodies [] and other physiologically active proteins produced by use of recombinant technology []. However, after early experiments, the use of this chromatographic technique for both analytical and preparative separation of proteins from human serum and plasma is relatively uncommon []. Moreover, despite its high potential, HIC is hardly ever used for sample preparation in order to reduce the complexity of biological materials prior to proteomic analyses [].…”

Section: Introductionmentioning

confidence: 99%

Separation of proteins from human plasma by sample displacement chromatography in hydrophobic interaction mode

et al. 2012

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Sample displacement chromatography (SDC) in reversed-phase and ion-exchange modes was introduced approximately twenty years ago. This method was first used for the preparative purification of peptides and proteins. Recently, SDC in ion-exchange mode was also successfully used for enrichment of low abundance proteins from human plasma. In this paper, the use of SDC for the separation of plasma proteins in hydrophobic interaction mode is demonstrated. By use of two or more columns coupled in series during sample application, and subsequent elution of detached columns in parallel, additional separation of bound proteins was achieved. Further low-abundance, physiologically active proteins could be highly enriched and detected by ESI-MS/MS.

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Hydrophobic Interaction Chromatography

Tomaz

Queiroz

2013

Liquid Chromatography

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Analysis of human α-thrombin by hydrophobic interaction high-performance liquid chromatography

Cited by 5 publications

References 13 publications

Comparison of two RP-HPLC methods for determination of recombinant human thrombin in pharmaceutical formulations. / Porovnanie dvoch RP-HPLC metód pre stanovenie rekombinantného ľudského trombínu vo farmaceutických formuláciách

Comparison of two RP-HPLC methods for determination of recombinant human thrombin in pharmaceutical formulations. / Porovnanie dvoch RP-HPLC metód pre stanovenie rekombinantného ľudského trombínu vo farmaceutických formuláciách

Separation of proteins from human plasma by sample displacement chromatography in hydrophobic interaction mode

Hydrophobic Interaction Chromatography

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